Anti RFP  

Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: HopERCre/+; R26Tom/+ mice. Fixation: 0.5% PFA. Antigen retrieval: Tamoxifen. Primary antibody: RFP antibody at 10 μg/mL for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: RFP is nuclear and occasionally cytoplasmic. Staining: Hop-derived cells in the hair follicle, labeled in red. Courtesy of Rajan Jain at UPenn.

Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: DsRed transgenic mouse retina. Fixation: 4% PFA. Blocking: 3% BSA, 0.3% Triton Primary antibody: RFP antibody at 1:100 for 12 h at 4°C. Secondary antibody: Alexa488 secondary antibody at 1:10,000 for 4 hours at RT. Localization: RFP is nuclear and occasionally cytoplasmic. Staining: labeled in red. Courtesy of Ismail Zaitoun at Wisc.edu.

Immunohistochemistry of Anti-RFP Antibody. Tissue: Mouse gut tissue in tomato transgenic mice. Fixation: formalin fixed paraffin embedded. Antigen retrieval: heat 5 min at high temp in 1X rodent decloaker. Primary antibody: RFP antibody at 1:200 for 1 h 30 min at RT. Secondary Antibody: HRP anti-rabbit (p/n 611-103-122). Image courtesy: Kwan Hyun Kim.

Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: (10X) Mouse lung tissue. Fixation: 4% PFA. Antigen retrieval: Heat. Primary antibody: Anti-RFP antibody at 1:50 for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:250 for 1 hr at RT. Staining: SPC+ cells are RFP positive in red. Courtesy of Johnathan C. Fox MD. PhD.

Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: (10X) Mouse E14.5 embryo heart tissue. Fixation: 4% PFA. Antigen retrieval: Heat. Primary antibody: Anti-RFP antibody at 1:50 for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:250 for 1 hr at RT. Staining: cardiac cells are RFP positive in red in tomato transgenic mice. Courtesy of Johnathan C. Fox MD. PhD.

Western Blot of RFP Antibody Pre-Absorbed. Lane 1: RFP (p/n 000-001-379). Lane 2: Human IgG (p/n 009-0102). Lane 3: Goat IgG (p/n 005-0102). Lane 4: Mouse IgG (p/n 010-0102). Load: 50ng per lane. Primary antibody: RFP Antibody Pre-Absorbed at 1:1,000 overnight at 4°C. Secondary antibody: Peroxidase conjugated rabbit secondary antibody (p/n 611-103-122) at 1:40,000 for 30 min at RT. Block: MB-070 Blocking Buffer for 30 min at RT. Predicted/Observed size: 27 kDa, 30 kDa.

Western Blot of Rabbit anti-RFP Antibody. Lane 1: Opal Prestained Marker (p/n MB-210-0500). Lane 2: 50ng of RFP. Primary Antibody: Anti-RFP at 1μg/mL overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit HRP (p/n 611-103-122) at 1:70,000 for 30mins at RT. Block: BlockOut Universal blocking buffer (p/n MB-073). Expect ~27kDa.

Western Blot of Rabbit anti-RFP antibody. Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 1: HEK293 lysate (p/n W09-000-365). Lane 2: HeLa Lysate (p/n W09-000-364). Lane 3: CHO/K1 Lysate (p/n W07-000-357). Lane 4: MDA-MB-231 (p/n W09-001-GK6). Lane 5: A431 Lysate (p/n W09-000-361). Lane 6: Jurkat Lysate (p/n W09-001-370). Lane 7: NIH/3T3 Lysate (p/n W10-000-358). Lane 8: E-coli HCP Control (p/n 000-001-J08). Lane 9: FLAG Positive Control Lysate (p/n W00-001-383). Lane 10: Red Fluorescent Protein (p/n 000-001-379). Lane 11: Green Fluorescent Protein (p/n 000-001-215). Lane 12: Glutathione-S-Transferase Protein. Lane 13: Maltose Binding Protein (p/n 000-001-385). Load: 10 μg of lysate or 50ng of purified protein per lane. Primary antibody: RFP antibody at 1ug/mL overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n 611-103-122) at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted/Observed size: 30 kDa for RFP.

Western Blot of Rabbit Anti-RFP MW Hu, Ms, Rt Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: RFP/HEK293T WCL (p/n 000-001-379/W09-001-GX5) [0.05/10μg] [+]. Lane 3: HEK293T Whole Cell Lysate [-]. Lane 4: RFP/NIH/3T3 Whole Cell Lysate (p/n 000-001-379/W10-000-358) [0.05/10μg] [+]. Lane 5: NIH/3T3 Whole Cell Lysate [-]. Lane 6: RFP/PC-12 Whole Cell Lysate (p/n 000-001-379/W12-001-GL9) [0.05/10μg] [+]. Lane 7: PC-12 Whole Cell Lysate [-]. Primary Antibody: Anti-RFP at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG Peroxidase (p/n 611-103-122) at 1:70,000 at RT for 30 mins. Predicted/Observed MW: ~27kDa. Exposure: 2 sec.

Western blot of RFP recombinant protein detected with Rockland's polyclonal anti-RFP antibody. Lane 1 shows no reaction against a GFP recombinant protein present in 10 μg of HeLa cell extract. Lane 2 shows a single band detected in 10 μg of a HeLa lysate containing RFP recombinant protein as a 27 kDa band. A 4-12% Bis-Tris gradient gel (Invitrogen) was used for SDS-PAGE. The membrane was blocked and then probed with Anti-RFP diluted 1:2,500 for 1 h at RT followed by washes and reaction with a 1:5,000 dilution of IRDye？800 conjugated Goat-a-Rabbit IgG [H&L] MX (611-132-122). IRDye？800 fluorescence image was captured using the OdysseyR Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.